Compositions comprising an antigen, an amphipathic compound and a hydrophobic carrier, and uses thereof
a technology of amphipathic compound and carrier, which is applied in the direction of antibody medical ingredients, drug compositions, immunological disorders, etc., can solve the problems of premature or accelerated release of antigens and other components, separation of aqueous and oily phases of composition,
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example 1
[0083]Pathogen free, female C57BL / 6 mice, 6-8 weeks of age, were obtained from Charles River Laboratories (St Constant, Quebec, Canada) and were housed according to institutional guideline with water and food ad libitum, under filter controlled air circulation.
[0084]The C3 cell line used in this study, a well-described mouse model for pre-clinical cervical cancer research, is an HPV 16-expressing C3 tumor cells derived from B6 mouse embryo cells (B6mec) and transformed with the complete HPV 16 genome under its own promoter and an activated-ras oncogene. The C3 cell line develops tumors when injected subcutaneously and has been used in cancer challenge studies to examine the efficacy of vaccine administered before or after C3 tumor cell implantation. The C3 cell line was maintained in Iscove Modified Dulbecco's Medium (IMDM; Sigma, St. Louis, Mo.) supplemented with 10% heat-inactivated fetal calf serum (Sigma, St. Louis, Mo.), 2 mM l-glutamine, 50 mM 2-mercaptoethanol, penicillin and...
example 2
[0092]Pathogen free, female C57BL / 6 mice, 6-8 weeks of age, were obtained from Charles River Laboratories (St Constant, Quebec, Canada) and were housed according to institutional guideline with water and food ad libitum, under filter controlled air circulation.
[0093]The HPV 16 E7 (H-2 Db) peptide RAHYNIVTF49-57 (SEQ ID NO: 1) containing a CTL epitope was fused to PADRE containing a CD4+ helper epitope by Dalton Chemical Laboratories Inc. (Toronto, Ontario, Canada). This peptide is hereafter designated FP and was used as antigen in vaccine at 20 micrograms / 100 microliters dose.
[0094]Vaccine efficacy was assessed by Enzyme-linked Immunospot assay (ELISPOT), a method that allows the ex vivo detection of antigen-specific cellular immune responses in splenocytes harvested from immunized C57BL / 6 mice. The ELISPOT assay is useful for assessing the presence / absence of an antigen-specific immune response but has its limitations when used as a correlate of vaccine efficacy against a target in...
example 3
[0100]In this example, poly IC double stranded RNA (Pierce, Milwaukee, USA) was used as a representative molecule that has physical and chemical characteristics that are similar to those of a genetic antigen construct (a nucleotide based plasmid or RNA molecule). PolyIC also serves as a representative of nucleotide based adjuvants that may be co-formulated with antigen in the invention. To formulate 1 ml final volume of vaccine described herein, 120.0 mg of dioleoyl phosphatidylcholine (DOPC) was solubilized in 480 ul of tert-butanol at a temperature of 40° C. for 10 to 15 minutes with shaking. polyI:C was first solubilized in water at a concentration of 5 mg / ml. Then 80 ul of polyI:C (0.4 mg) were further diluted in 320 ul of water. The polyI:C dilution was then added and mixed to the DOPC / tert-butanol mixture in vial 21. A dry homogenous mixture of DOPC / adjuvant was prepared by removing the solvent and water present in the formulation by lyophilization. A control formulation conta...
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